Regulation of nuclear factor-kappa B and activator protein-1 activities after stimulation of T cells via glycosylphosphatidylinositol-anchored Ly-6A/E

J Immunol. 1994 Sep 15;153(6):2394-406.


Cross-linking of glycosylphosphatidylinositol-anchored proteins, including mouse Ly-6A/E, leads to IL-2 secretion and T cell activation, whereas engagement of Ly-6A/E uniquely inhibits IL-2 production induced via TCR. However, little is known concerning the molecular mechanism by which glycosylphosphatidylinositol-anchored proteins regulate IL-2 expression. In this study, we have examined the ability of an anti-Ly-6A/E mAb to regulate transcription factors controlling IL-2 expression. Stimulation of EL4J(Ly-6E).A4 cells with anti-CD3 epsilon or anti-Ly6A/E mAbs induced nuclear factor (NF)-kappa B p65-p50 (RelA/p50) and AP-1 (Fos/Jun) binding activities and increased nuclear factor of activated T cells (NF-AT) activity, whereas octamer-binding factor and NF-Y levels were stable. Cyclic AMP response element binding protein and T cell-specific factor-1 (alpha) activities were selectively enhanced by anti-CD3 epsilon, but not by anti-Ly6A/E, which suggests that signaling via the TCR and Ly-6 were not identical. Costimulation of these cells with both mAbs produced substantially reduced levels of AP-1, NF-AT, and, especially, NF-kappa B p65-p50 whereas cyclic AMP response element binding protein and T cell-specific factor-1(alpha) were induced to a level seen after stimulation by anti-CD3 epsilon. The inducibility of the IL-2 enhancer in vivo and the contribution of individual transcription factors for this induction were assessed with use of reporter chloramphenicol acetyltransferase constructs containing the IL-2 enhancer or oligomerized binding sites for transcription factors. These experiments also demonstrated a key role for NF-kappa B and AP-1 in the transcriptional regulation of the IL-2 gene by TCR- and Ly6A/E-mediated signaling. By using the 2B4.11 T cell hybridoma and a mutated variant, were revealed a crucial role for the zeta-chain in Ly6A/E-mediated activation of NF-kappa B.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies, Monoclonal
  • Antigens, Ly / immunology*
  • Antigens, Surface / immunology
  • Base Sequence
  • CD3 Complex / immunology
  • Cell Line
  • Glycosylphosphatidylinositols / metabolism
  • Hybridomas
  • Interleukin-2
  • Membrane Glycoproteins / immunology
  • Membrane Proteins / immunology*
  • Molecular Sequence Data
  • NF-kappa B / metabolism*
  • Proto-Oncogene Proteins c-jun / metabolism*
  • Receptors, Antigen, T-Cell / immunology
  • Signal Transduction / physiology
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism
  • Thy-1 Antigens
  • Transcription Factors / metabolism
  • Transfection


  • Antibodies, Monoclonal
  • Antigens, Ly
  • Antigens, Surface
  • CD3 Complex
  • Glycosylphosphatidylinositols
  • Interleukin-2
  • Ly6a protein, mouse
  • Membrane Glycoproteins
  • Membrane Proteins
  • NF-kappa B
  • Proto-Oncogene Proteins c-jun
  • Receptors, Antigen, T-Cell
  • Thy-1 Antigens
  • Transcription Factors
  • antigen T cell receptor, zeta chain